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Biomedical subjects

H Kubo

Publications and source records attributed to H Kubo.

At least 289 records · Page 16Linked to original sources

A new enzymatic cycling technique for glutamate determination in brain microdialysates.

A quantitative analysis of glutamate in brain dialysate was made by using an enzymatic cycling technique. This method made it possible to measure the concentration of glutamate in dialysate collected at 30-s intervals. Dialysates were collected from Mongolian gerbil hippocampus before, during, and after two 90-s ischemic insults at an interval of 5 min. An extracellular increase in levels of glutamate was already observed in samples collected during a 30-60 s period after the onset of each ischemia, and the levels of glutamate were maximal at the end of each period of ischemia (approximately a fourfold increase). The increased levels of glutamate rapidly returned almost to preischemic levels by 30 s of recirculation. This method will provide more precise information about temporal changes in the extracellular glutamate concentration in the brain during ischemia.

Animals↗

Immunogold-silver staining method for light and electron microscopic detection of lymphocyte cell surface antigens with monoclonal antibodies.

We used the immunogold-silver staining method (IGSS) for detection of lymphocyte cell surface antigens with monoclonal antibodies in light and electron microscopy and compared this procedure with the immunogold staining method. Two different sizes of colloidal gold particles (5 nm and 15 nm) were used in this study. Immunolabeling on cell surfaces was visualized as fine granules only by IGSS in light microscopy. The labeling density (silver-gold complexes/cell) and diameters of silver-enhanced gold particles on cell surfaces were examined by electron microscopy. Labeling density was influenced not by the enhancement time of the physical developer but by the size of the gold particles. However, the development of shells of silver-enhanced gold particles correlated with the enhancement time of the physical developer rather than the size of the colloidal gold particles. Five-nm gold particles enhanced with the physical developer for 3 min were considered optimal for this IGSS method because of reduced background staining and high specific staining in the cell suspensions in sheep lymph. Moreover, this method may make it possible to show the ultrastructure of identical positive cells detected in 1-micron sections counterstained with toluidine blue by electron microscopy, in addition to the percentage of positive cells by light microscopy.

Animals↗

Liposomes prepared from synthetic amphiphiles. II. Their interaction with Ehrlich ascites tumor cells and tissue distribution in Ehrlich solid tumor-bearing mice.

The interaction of 99mTc-labeled liposomes prepared from synthetic amphiphiles containing amino acid residues with Ehrlich ascites tumor cells in vitro and their tissue distributions in Ehrlich solid tumor-bearing mice were investigated. The amphiphiles used were N,N-didodecyl-N alpha-[6-(trimethylammonio)hexanoyl]-L-alaninamide bromide N+C5Ala2C12), N,N-didodecyl-N alpha-(6-[dimethyl(2-carboxyethyl)ammonio]hexanoyl)-L- alaninamide bromide (CAC2N+C5Ala2C12) and S-[1-carboxy-2- ([2,3-bis(hexadecyloxy)propoxy]carbonyl)ethyl]homocysteine (HcyM-G2C1 6). Most of the radioactivity of N+C5Ala2C12 and CAC2N+C5Ala2C12 liposomes was firmly bound to Ehrlich ascites tumor cells in vitro. On the other hand, the accumulation of three 99mTc-labeled liposomes in the tumor of Ehrlich solid tumor-bearing mice was low (about 1% dose per gram of tissue), and most of the liposomes were taken up highly in the liver and spleen of the tumor-bearing mice. However, the radioactivity of the liposomes in the tumor, especially that of N+C5Ala2C12 and CAC2N+C5Ala2C12 liposomes, decreased more slowly with time than in the liver in up to 24 h after injection, suggesting that these liposomes were hard to separate from the tumor cells.

Animals↗

Immunohistochemical differentiation between lymphatic vessels and blood vessels--use of anti-basement membrane antibodies and anti-factor VIII-related antigen.

Several immunohistochemical methods using Factor VIII-Related antigen (FVIIIR:Ag), laminin, Type IV collagen and fibronectin antisera were applied for the purpose of differentiating rat lymphatics from blood vessels by light and electron microscopy. Weibel-Palade bodies (WPB) were demonstrated in both types of vessels by conventional electron microscopy. The immunoreactivity to laminin and Type IV collagen in blood vessels showed a strong, continuous, linear subendothelial staining pattern in contrast to lymphatic vessels in which immunoreactivity was absent or weak in paraffin-embedded sections stained with the indirect immunoperoxidase technique. A positive reaction for fibronectin was observed in all extra-vascular tissue spaces as well as in lymphatics and blood vessels. FVIIIR:Ag and WPB were present in both lymphatic and blood endothelial cells. FVIIIR:Ag antiserum labeled with gold particles was observed only in the vacuoles which were assumed to be identical with WPB as demonstrated by our conventional electron microscopy. We conclude that the immunohistochemical method using laminin and Type IV collagen antisera is a reliable and practical way to differentiate lymphatic vessels from blood vessels by light microscopy.

Animals↗

[Efficacy of combined therapy using coronary reperfusion and elective percutaneous transluminal coronary angioplasty for acute myocardial infarction].

The effects of elective percutaneous transluminal coronary angioplasty (PTCA) performed one month after coronary reperfusion therapy in patients with acute myocardial infarction (AMI) were observed using exercise Tl-201 myocardial scintigraphy performed before and after PTCA. Myocardial perfusion in Tl-201 scintigraphy was significantly greater in the early (less than 4 hours) and late (4-9 hours) reperfusion groups than in the total occlusion group one month after the onset of AMI. Both reperfusion groups showed significant improvement in myocardial perfusion after elective PTCA; whereas, the total occlusion group showed no significant improvement. In the early reperfusion group, there were no significant differences in myocardial perfusion between those with well developed and those with poorly developed collateral circulations one month after the onset of AMI. However, in the late reperfusion group, myocardial perfusion was greater in those with well developed collateral circulations compared to those with poorly developed collateral circulations. The grade of myocardial perfusion in the late reperfusion group with poorly developed collateral circulations did not differ significantly from that of the total occlusion group. There was significant improvement of myocardial perfusion in the early and late reperfusion groups with well developed collateral circulations after elective PTCA; whereas, no significant improvement was observed in the late reperfusion group with poorly developed collateral circulations. These findings indicate that the time interval from the onset of AMI to reperfusion and the grade of development of collateral circulations are the major determinants of myocardial perfusion after elective PTCA and after reperfusion therapy.

Adult↗

[Serum beta 2-microglobulin concentration and its removal in patients treated with continuous ambulatory peritoneal dialysis].

This study was performed to clarify serum beta-2 microglobulin (beta 2-M) level and its change in 50 CAPD and 56 HD patients. There was significant correlation between duration of dialysis and serum beta 2-M level in CAPD and HD patients treated under 12 months, but no correlation in those treated over 12 months. Serum beta 2-M level was 33.5 +/- 9.1 mg/l in 45 CAPD patients treated over 12 months, and 46.2 +/- 21.1 mg/l in 35 HD patients. In 26 CAPD patients treated over 12 months, clearance and removal of beta 2-M were 1.0 +/- 0.3 ml/min and 43.0 +/- 17.8 mg/day. There was significant correlation between dwell time and beta 2-M removal (p less than 0.01), and these results suggested beta 2-M was removed by diffusion. Because CAPD treatment can lower serum beta 2-M level compared to HD, there is possibility that CAPD is useful at prevention of dialysis associated amyloidosis.

Adult↗

Increase in lens gangliosides due to aging and cataract progression in human senile cataract.

Gangliosides were isolated from human senile cataractous lenses by solvent extraction, DEAE-Sephadex column chromatography, and thin-layer chromatography. The content and composition of gangliosides were examined in individual lens tissues. Three predominant gangliosides, GM3, GM1, and GD1a, were tentatively identified in comparison with authentic brain gangliosides, and several unidentified gangliosides were also recognized. The increase in ganglioside content per mg of protein content in cataractous lenses was found to be influenced by two physiologic parameters: aging and cataract progression. The mature cataractous lenses showed a higher ganglioside level on a protein basis than the immature lenses compared with the same age group. On the basis of statistical analysis, an age-dependent increase in ganglioside concentration was recognized in both mature and immature lens groups. The relative increase in slow-moving polysialogangliosides on thin-layer chromatography seemed to be caused by the maturation of cataract. The sugar composition of one of the polysialogangliosides was found to be glucose, galactose, and sialic acid in the molar ratio of 2:1:4; this suggests the presence of a unique ganglioside species in human cataractous lens.

Adult↗

[Activated pulmonary intravascular macrophages increase pulmonary microvascular permeability in sheep].

Pulmonary intravascular macrophages (PIMs) reside in the alveolar capillaries of sheep lung. However, the role of PIMs activation in the mechanism of increasing pulmonary microvascular permeability is not clear. We determined, using awake sheep with chronic lung lymph fistulas, the effects of intraarterial infusion of latex beads emulsion (1 microns in diameter) on lung lymph-dynamics. Sheep were divided into 3 groups; control, low dose latex and high dose latex groups. In control group, their lymph-dynamics were stable during the experimental period. In low dose latex group (5.46 x 10(9) beads/kg), lung lymph flow increased during the infusion period, but lung lymph protein clearance did not change. In high dose latex group (5.46 x 10(10) beads/kg), lung lymph flow began to increase during the infusion period and remained high, and lung lymph protein clearance increased significantly during and after the infusion of latex beads emulsion. The latex beads mainly distributed to lungs, and 70% of the beads were caught by PIMs in their phagosomes. These findings suggest that the activation of PIMs through their phagocytic uptake of circulating microparticles increases pulmonary microvascular permeability.

Animals↗

Lymphatics, intraepithelial lymphocytes and endometrial lymphoid tissues in the rabbit uterus: an electron microscopic and immunohistological study.

Rabbit uterine intraepithelial lymphocytes, endometrial lymphoid aggregates and lymphatic capillaries were examined electron-microscopically and immunohistochemically at well-defined intervals after the injection of human chorionic gonadotropin (hCG). Lymphatic capillaries originated near the bases of the glands in the uterine cervix and in the border zone between the lamina propria mucosae and myometrium in the uterine body. The lymphatic capillaries were maximally dilated, and their endothelial cells were thinnest 8 hours after hCG injection. Patent junctions between the adjacent endothelial cells of lymphatics in the uterine body were observed in good accordance with the appearance of stromal edema and lymphatic dilatation. The numbers of intraepithelial lymphocytes and macrophages changed cyclically after the induction of ovulation. They were highest 11 hours after hCG injection when lymphocytes were seen occasionally in the lumens of lymphatics located in the lamina propria mucosae of the uterine body. Most of the intraepithelial and interstitial lymphocytes were cells labeled with T cell serum but some were labeled with IgA serum and were occasionally seen beneath the epithelium. Lymphoid aggregates were uniformly present in the stratum basalis and consisted of lymphocytes and macrophages. They had no germinal centers, surrounding lymphatics or high endothelial venules (HEV). The results suggest that lymphatic capillaries are the main route for the removal of edema fluid and for migratory lymphoid cells in the rabbit uterus.

Animals↗

[Clinical reevaluation of continuous intravenous infusion of 5-fluorouracil--plasma concentrations and clinical dose by continuous intravenous and 60-min infusions].

Daily and intermittent continuous intravenous infusions [by gravity drip, (IVG) or infusion pump, (IVP)] and intermittent short-time intravenous drip infusion of 5-FU were carried out on advanced cancer patients. The MTD and dose-limiting toxicity were investigated in relation to the plasma concentrations of 5-FU determined by HPLC. Responses in eleven patients receiving IVG administration daily at 8-21 mg/kg/day were NC, but those given 5-FU alone showed no adverse reactions. Plasma concentrations were too low to be determined. In 9 patients receiving IVG or IVP administration weekly at 60 mg/kg for 24 hr, 1 of the 5 evaluable patients showed reduced hepatic metastatic lesions. One of 4 patients receiving IVP administration weekly at 120 mg/kg for 48 hr showed a disappearance of metastatic lesions in the skeletal muscle, but bone marrow suppression was observed as dose-limiting toxicity. Pharmacokinetics were more stable in IVP than in IVG with less individual difference in the plasma concentrations. Among the outpatients receiving short-time iv, IVG administration once or twice a week, 2 patients given weekly administrations at 20 mg/kg for 60 min showed slight adverse reactions. In 6 patients given high-dose administrations, bone marrow suppression was observed. When pharmacokinetics in the patients given 5-FU for 60 min were compared between the IVG and IVP groups, there were individual differences in plasma concentrations, but the differences were not significant. It was concluded from above results that the following practical dose schedules would be recommendable: 60 mg/kg for 24hr/week by IVP for inpatients and 20 mg/kg for 60 min/week by IVG for outpatients.

Adult↗

[A case of primary low spinal fluid pressure syndrome complicated by bilateral subdural hematoma--study of the cerebrospinal fluid circulation].

A 43-year-old man with no history of trauma or neurosurgical procedure was admitted our hospital because of orthostatic headache made worse in upright position. Spinal fluid pressure was 0 mmH2O, and CT scan revealed wide cortical sulci and ventricles. In the 26th hospital day he had severe headache and CT scan revealed bilateral subdural hematoma, then neurosurgical procedure was done. About 2 months later, he was relieved of the headache and spinal fluid pressure returned to normal. We performed lumbar isotope cisternography and isotope choroid plexography in our case when low spinal fluid pressure and normalized pressure. Isotope cisternography revealed the radioactivity counts decreased rapidly in low pressure and normal in normal pressure. On the other hand isotope choroid plexography was almost no difference between low and normal pressure. Our data suggest that primary low spinal fluid pressure syndrome is not caused by hypoproduction, but by an undetectable leak from a minute tear in a spinal root sleeve or hyperabsorption at the spinal arachnoid membrane.

Adult↗

[Three cases of pneumothorax due to lung cancer].

Spontaneous pneumothorax due to primary lung cancers are increasingly reported as of lung cancer incidence increased. We experienced one lung cancer case with pneumothorax as a primary symptom and two who developed pneumothorax after chemotherapy, among 238 cases of primary lung cancer. In the literature, 76 cases of pneumothorax due to primary lung cancers are described. The mechanism of pneumothorax is discussed.

Adult↗

Structural parts involved in activation and inactivation of the sodium channel.

Structure-function relationships of the sodium channel expressed in Xenopus oocytes have been investigated by the combined use of site-directed mutagenesis and patch-clamp recording. This study provides evidence that the positive charges in segment S4 are involved in the voltage-sensing mechanism for activation of the channel and that the region between repeats III and IV is important for its inactivation.

Action Potentials↗

Usefulness of anaerobic threshold in estimating intensity of exercise for diabetics.

We examined the utility of the anaerobic threshold (AT) for quantifying the intensity of exercise that a diabetic patient is capable of handling. Thirteen diabetic patients treated with buformin exercised on a bicycle ergometer, and comparison was made with 20 healthy subjects matched for age and sex. The AT was determined from VO2 and VE with a personal computer. The intensity of exercise at the AT was 93 +/- 6 W in diabetic men and 80 +/- 10 W in diabetic women, values that were less than those of healthy subjects (P less than 0.05). There was a negative correlation between the intensity of exercise at the AT and the plasma concentration of buformin (P less than 0.01). There were no significant differences in either plasma lactic acid or pyruvic acid concentration at the AT between healthy subjects and diabetics. The plasma glucose at the AT or after exercise was lower than the baseline values in all subjects (P less than 0.01). The plasma insulin at the AT was lower than the baseline values in healthy subjects (P less than 0.01), but not in diabetics. There were no changes in plasma glucagon in any group. We concluded that determination of the AT is a simple, non-invasive procedure useful for ascertaining the optimal intensity of exercise for diabetics.

Adult↗

Thromboxane A2 synthetase inhibitors. 2. Syntheses and activities of tetrahydronaphthalene and indan derivatives.

A series of 1-imidazolylalkyl-substituted or 5-thiazolylalkyl-substituted tetrahydronaphthalenecarboxylic acid and indancarboxylic acid derivatives were prepared and tested for the inhibitory activities of thromboxane A2 (TXA2) production in vitro and ex vivo. Most of the compounds showed potent TXA2 synthetase inhibitory activities in vitro and had long duration of inhibition of TXA2 production in rats when orally or intravenously administrated. The imidazole analogues had slightly less potency in vitro than the thiazole analogues, but the activities of the imidazole analogues in ex vivo models were equal or superior to the activities of the thiazole analogues. 6-(1-Imidazolyl-methyl)-5,6,7,8-tetrahydronaphthalene-2-carboxylic acid hydrochloride hemihydrate (47a, DP-1904) was chosen for clinical studies.

6-Ketoprostaglandin F1 alpha↗